Systems, methods, and apparatus for increased wastewater effluent and biosolids quality

ABSTRACT

Methods of delivering microorganisms loaded onto an inorganic porous medium. Methods of treating wastewater to increase effluent and biosolids quality. Methods of reducing ammonia and denitrifying wastewater effluent. Methods of reducing phosphorous concentration in wastewater effluent. Composition of biosolids derived from wastewater treatment. Wastewater treatment assemblage for increasing wastewater effluent and biosolids quality.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.16/489,310, filed Aug. 27, 2019, which is a continuation ofInternational Application No. PCT/US2018/019961, filed Feb. 27, 2018,which claims the benefit of priority to U.S. Provisional Application No.62/464,816, filed on Feb. 28, 2017, all of which are incorporated hereinby reference.

SUMMARY OF THE INVENTION

Disclosed herein, in certain embodiments, are methods of producing afertilizer or compost comprising: (a) proving a microbial solutioncomprising at least one microbial species loaded onto an inorganicporous medium; (b) providing a reaction vessel comprising an influentstream, an effluent stream, an aqueous phase, and a biosolids phase andwherein the biosolids phase comprises at least one nutrient source forthe at least one microbial species; (c) adding the microbial solutioncomprising at least one microbial species loaded onto the inorganicporous medium to the reaction vessel and wherein the at least onemicrobial species consumes a portion of the biosolids phase; (d)separating the effluent stream into a treated aqueous phase and atreated biosolids phase; and (e) dewatering the treated biosolids phaseto produce fertilizer or compost. In some embodiments, the inorganicporous medium comprises silica. In some embodiments, the inorganicporous medium comprises zeolite. In some embodiments, the inorganicporous medium comprises silica, zeolite, aluminosilicate, silicate,diatomaceous earth, or any combination thereof. In some embodiments, theat least one microbial species is aerobic. In some embodiments, the atleast one microbial species is anaerobic. In some embodiments, the atleast one microbial species is facultative.

In some embodiments, the influent stream includes residentialwastewater, agricultural wastewater, industrial wastewater, runoffwastewater, or any combination thereof. In some embodiments, adding themicrobial solution comprising the at least one microbial species loadedonto the inorganic porous medium to the reaction vessel reduces thequantity of the treated biosolids phase. In some embodiments, adding themicrobial solution comprising the at least one microbial species loadedinto the inorganic porous medium to the reaction vessel increases thecarrying capacity of the reaction vessel. In some embodiments,separating the effluent stream into the treated aqueous phase and thetreated biosolids phase includes decantation, filtration,centrifugation, or any combination thereof. In some embodiments,dewatering the treated biosolids phase includes centrifugation orfiltration.

In some embodiments, the treated biosolids phase comprises less thanabout 3 most probable number (MPN) Salmonella enterica spp. per 4 gramsof solids on a dry basis. In some embodiments, the treated biosolidsphase comprises less than about 1000 MPN fecal coliform per gram oftotal solids on a dry basis. In some embodiments, the treated biosolidsphase comprises less than about 1 plaque-forming unit (PFU) of entericviruses per 4 grams of total solids on a dry basis. In some embodiments,the treated biosolids phase comprises less than about 1 viable helminthova per 4 grams of total solids on a dry basis with a vector attractionstandard oxygen uptake rate of less than about 1.5 milligrams of oxygenper gram of solids per hour. In some embodiments, the treated biosolidsphase comprises less than about 41 ppm arsenic, less than about 39 ppmcadmium, less than about 1,200 ppm chromium, less than about 1,500 ppmcopper, less than about 300 ppm lead, less than about 17 ppm mercury,less than about 420 ppm nickel, less than about 36 ppm selenium, andless than about 2,800 ppm zinc. In some embodiments, the inorganicporous medium is selected or modified for sorption of nitrogen and/orphosphorous from the aqueous phase to (i) increase a nutrientconcentration of the fertilizer of compost and (ii) decrease the amountof nitrogen and/or phosphorous in the aqueous phase. In someembodiments, (i) a solids retention time in the reaction vessel isincreased by greater than or equal to about 50% and/or (ii) an amount ofthe treated biosolids phase is reduced by greater than or equal to about5% when the solids retention time is maintained constant. In someembodiments, a fertilizer or compost is produced by the method.

Disclosed herein, in certain embodiments, are methods of producingfertilizer or compost from a wastewater treatment plant, the methodcomprising: (a) providing a microbial solution comprising at least onemicrobial species loaded onto an inorganic porous medium; (b) providingan aeration basin comprising an influent stream, an effluent stream, anaqueous phase, and a biosolids phase and wherein the biosolids phasecomprises at least one nutrient source for the at least one microbialspecies; (c) adding the microbial solution comprising the at least onemicrobial species loaded onto the inorganic porous medium to theaeration basin and wherein the at least one microbial species consumes aportion of the biosolids phase; (d) separating the effluent stream intoa treated aqueous phase and a treated biosolids phase; (e) returning aquantity of the treated biosolids phase to the aeration basin, whereinthe treated biosolids phase is further consumed by the at least onemicrobial species and wherein the quantity of the treated biosolidsphase is reduced; (f) digesting the treated biosolids phase in adigester to yield a digested biosolids phase; and (g) dewatering thedigested biosolids phase to produce a fertilizer or compost. In someembodiments, the inorganic porous medium comprises silica. In someembodiments, the inorganic porous medium comprises zeolite. In someembodiments, the inorganic porous medium comprises silica, zeolite,aluminosilicate, silicate, diatomaceous earth, or any combinationthereof. In some embodiments, the at least one microbial species isaerobic. In some embodiments, the at least one microbial species isanaerobic. In some embodiments, the at least one microbial species isfacultative.

In some embodiments, the influent stream includes residentialwastewater, agricultural wastewater, industrial wastewater, runoffwastewater, or any combination thereof. In some embodiments, adding themicrobial solution comprising the at least one microbial species loadedonto the inorganic porous medium to the aeration basin reduces thequantity of the treated biosolids phase. In some embodiments, adding themicrobial solution comprising the at least one microbial species loadedonto the inorganic porous medium to the aeration basin increases thecarrying capacity of the aeration basin. In some embodiments, separatingthe effluent stream into the treated aqueous phase and the treatedbiosolids phase includes decantation, filtration, centrifugation, or anycombination thereof. In some embodiments, separating the effluent streaminto the treated aqueous phase and the treated biosolids phase includesdecantation, filtration, centrifugation, or any combination thereof. Insome embodiments, dewatering the digested biosolids phase includescentrifugation or filtration.

In some embodiments, the digested biosolids phase comprises less thanabout 3 most probable number (MPN) Salmonella enterica spp. per 4 gramsof solids on a dry basis. In some embodiments, the digested biosolidsphase comprises less than about 1000 MPN fecal coliform per gram oftotal solids on a dry basis. In some embodiments, the digested biosolidsphase comprises less than about 1 plaque-forming unit (PFU) of entericviruses per 4 grams of total solids on a dry basis. In some embodiments,the digested biosolids phase comprises less than about 1 viable helminthova per 4 grams of total solids on a dry basis with a vector attractionstandard oxygen uptake rate of less than about 1.5 milligrams of oxygenper gram of solids per hour. In some embodiments, the digested biosolidsphase comprises less than about 41 ppm arsenic, less than about 39 ppmcadmium, less than about 1,200 ppm chromium, less than about 1,500 ppmcopper, less than about 300 ppm lead, less than about 17 ppm mercury,less than about 420 ppm nickel, less than about 36 ppm selenium, andless than about 2,800 ppm zinc. In some embodiments, the inorganicporous medium is selected or modified for sorption of nitrogen and/orphosphorous from the aqueous phase to (i) increase a nutrientconcentration of the fertilizer of compost and (ii) decrease the amountof nitrogen and/or phosphorous in the aqueous phase. In someembodiments, (i) a solids retention time in the reaction vessel isincreased by greater than or equal to about 50% and/or (ii) an amount ofthe treated biosolids phase is reduced by greater than or equal to about5% when the solids retention time is maintained constant. In someembodiments, a fertilizer or compost is produced by the method. In someembodiments, a wastewater treatment facility implements the method.

Disclosed herein, in certain embodiments, are compositions of solidfertilizer or compost comprising: (i) dewatered biosolids, (ii) at leastabout 500 ppm of an inorganic porous medium on a dry basis; and whereinthe composition is characterized as having at least one of the followingfive properties: an analytical composition comprising less than about 41ppm arsenic, less than about 39 ppm cadmium, less than about 1,200 ppmchromium, less than about 1,500 ppm copper, less than about 300 ppmlead, less than about 17 ppm mercury, less than about 420 ppm nickel,less than about 36 ppm selenium, and less than about 2,800 ppm zinc; aconcentration of Salmonella enterica spp. below about 3 most probablenumber (MPN) per 4 grams of total solids on a dry basis; a totalconcentration of fecal coliform bacteria less than about 1000 MPN pergram of total solids on a dry basis; a density of enteric viruses lessthan about 1 plaque-forming unit (PFU) per 4 grams of total solids on adry basis; or a density of viable helminth ova less than 1 per 4 gramsof total solids on a dry basis with a vector attraction standard oxygenuptake rate of less than 1.5 milligrams of oxygen per gram of solids perhour. In some embodiments, the biosolids are not derived from awastewater treatment plant. In some embodiments the biosolids arederived from a wastewater treatment plant.

In some embodiments, the inorganic porous medium comprises silica. Insome embodiments, the inorganic porous medium comprises zeolite. In someembodiments, the inorganic porous medium comprises silica, zeolite,aluminosilicate, silicate, diatomaceous earth, or any combinationthereof. In some embodiments, the enteric viruses include humanastroviruses, human adenoviruses, noroviruses, human sapoviruses, humanparvoviruses, non-polio enteroviruses, and human rotaviruses. In someembodiments, the composition comprises an analytical compositioncomprising less than about 41 ppm arsenic, less than about 39 ppmcadmium, less than about 1,200 ppm chromium, less than about 1,500 ppmcopper, less than about 300 ppm lead, less than about 17 ppm mercury,less than about 420 ppm nickel, less than about 36 ppm selenium, andless than about 2,800 ppm zinc, a concentration of Salmonella entericaspp. below about 3 most probable number (MPN) per 4 grams of totalsolids on a dry basis, a total concentration of fecal coliform bacterialess than about 1000 MPN per gram of total solids on a dry basis, adensity of enteric viruses less than about 1 plaque-forming unit (PFU)per 4 grams of total solids on a dry basis, and a density of viablehelminth ova is less than about 1 per 4 grams of total solids on a drybasis with a vector attraction standard oxygen uptake rate of less than1.5 milligrams of oxygen per gram of solids per hour. In someembodiments, a wastewater treatment facility produces the composition.

Disclosed herein, in certain embodiments, are methods of reducingammonia and/or denitrifying wastewater comprising: (a) providing amicrobial solution comprising at least one microbial species loaded onan inorganic porous medium; (b) providing an aeration basis comprisingan influent stream, and effluent stream, an aqueous phase, and whereinthe aqueous phase comprises ammonia; and (c) adding the microbialsolution comprising the at least one microbial species loaded onto theinorganic porous medium to the aeration basin and wherein the at leastone microbial species consumes the ammonia to produce nitrite, nitrate,molecular nitrogen, or any combination thereof and thereby reduces theamount ammonia and/or denitrifies the wastewater. In some embodiments,the inorganic porous medium comprises silica. In some embodiments, theinorganic porous medium comprises zeolite. In some embodiments, theinorganic porous medium comprises silica, zeolite, aluminosilicate,silicate, diatomaceous earth, or any combination thereof.

In some embodiments, the at least one microbial species is aerobic. Insome embodiments, the at least one microbial species is anaerobic. Insome embodiments, the at least one microbial species is facultative. Insome embodiments, reducing ammonia does not require the use of achlorinator, ozone, peroxide, bleach, or ultra violet light. In someembodiments, the inorganic porous medium is selected or modified forsorption of nitrogen from the aqueous phase to (i) increase a nutrientconcentration of a solid phase and (ii) decrease the amount of nitrogenin the aqueous phase. In some embodiments, an effluent is produced usingthe methods. In some embodiments, a wastewater treatment facilityimplementing the method.

Disclosed herein, in certain embodiments, are methods of reducingphosphorous in wastewater comprising: (a) providing a microbial solutioncomprising at least one microbial species loaded onto an inorganicporous medium; (b) providing an aeration basin comprising an influentstream, an effluent stream, and an aqueous phase, and wherein theaqueous phase comprises inorganic and organic aqueous phosphorus; and(c) adding the microbial solution comprising the at least one microbialspecies loaded onto the inorganic porous medium to the aeration basinand wherein the at least one microbial species consumes the inorganicand organic aqueous phosphorus to reduce the amount of inorganic andorganic aqueous phosphorous and prevent eutrophication. In someembodiments, the inorganic porous medium comprises silica. In someembodiments, the inorganic porous medium comprises zeolite. In someembodiments, the inorganic porous medium comprises silica, zeolite,aluminosilicate, silicate, diatomaceous earth, or any combinationthereof.

In some embodiments, the at least one microbial species is aerobic. Insome embodiments, the at least one microbial species is anaerobic. Insome embodiments, the at least one microbial species is facultative. Insome embodiments, reducing the amount of inorganic and organic aqueousphosphorous does not require the addition of an iron or aluminacompound. In some embodiments, reducing the amount of inorganic andorganic aqueous phosphorous does not require the addition of magnesiumchloride or magnesium hydroxide. In some embodiments, the inorganicporous medium is selected or modified for sorption of the inorganic andorganic aqueous phosphorous from the aqueous phase to (i) increase anutrient concentration of a solid phase and (ii) decrease the amount ofinorganic or organic aqueous phosphorous in the aqueous phase. In someembodiments, an effluent is produced by the methods. In someembodiments, a wastewater treatment facility implements the methods.

Disclosed herein, in certain embodiments, are assemblages for treatingwastewater comprising: an influent stream comprising an aqueous phaseand a biosolids phase; a reaction vessel comprising at least one inletstream, at least one outlet stream, and at least one microbial speciesloaded onto an inorganic porous medium; and a separator comprising atleast one inlet stream and at least one outlet stream and wherein theseparator separates the aqueous phase from the biosolids phase. In someembodiments, the inorganic porous medium comprises silica. In someembodiments, the inorganic porous medium comprises zeolite. In someembodiments, the inorganic porous medium comprises silica, zeolite,aluminosilicate, silicate, diatomaceous earth, or any combinationthereof.

In some embodiments, the at least one microbial species is aerobic. Insome embodiments, the at least one microbial species is anaerobic. Insome embodiments, the at least one microbial species is facultative. Insome embodiments, the assemblage does not comprise a chlorinator. Insome embodiments, the assemblage does not comprise an ozonator. In someembodiments, the influent stream includes residential wastewater,agricultural wastewater, industrial wastewater, runoff wastewater, orany combination thereof. In some embodiments, adding the microbialsolution comprising the at least one microbial species to the reactionvessel reduces the quantity of the biosolids phase. In some embodiments,the separator comprises a decanter, filter, centrifuge, or combinationthereof. In some embodiments, the reaction vessel comprises an aerationbasin, lagoon, oxidation ditch, extended aeration, conventionalactivated sludge, membrane bioreactor, moving bed biofilm reactor,integrated fixed film activated sludge, trickle bed reactor, sequencingbatch reactor, complete mix, step feed, modified aeration, contactstabilization, high purity oxygen reactor, Kraus process, or any otherreactor for microbial growth. In some embodiments, the reaction vesselis an aeration basin. In some embodiments, a wastewater treatmentfacility comprises the assemblage.

Disclosed herein, in certain embodiments, are assemblages for treatingwastewater comprising: an influent stream comprising an aqueous phaseand a biosolids phase; a reaction vessel comprising at least one inletstream, at least one outlet stream, and at least one microbial speciesloaded onto an inorganic porous medium; a separator comprising at leastone inlet stream and at least one outlet stream and wherein theseparator separates the aqueous phase from the biosolids phase; a returnactivated sludge (RAS) line that carries a portion of the biosolidsphase from the separator to the aeration basin; a waste activated sludge(WAS) line that carries a second portion of the biosolids phase to adewaterer that dewaters the biosolids phase; and an effluent stream thatcomprises a treated aqueous phase. In some embodiments, the inorganicporous medium comprises silica. In some embodiments, the inorganicporous medium comprises zeolite. In some embodiments, the inorganicporous medium comprises silica, zeolite, aluminosilicate, silicate,diatomaceous earth, or any combination thereof.

In some embodiments, the at least one microbial species is aerobic. Insome embodiments, the at least one microbial species is anaerobic. Insome embodiments, the at least one microbial species is facultative. Insome embodiments, the assemblage does not comprise a chlorinator. Insome embodiments, the assemblage does not comprise an ozonator. In someembodiments, the influent stream includes residential wastewater,agricultural wastewater, industrial wastewater, runoff wastewater, orany combination thereof. In some embodiments, adding the microbialsolution comprising the at least one microbial species loaded onto aninorganic porous medium to the reaction vessel reduces the quantity ofthe biosolids phase. In some embodiments, the separator comprises adecanter, filter, centrifuge, or combination thereof. In someembodiments, the reaction vessel comprises an aeration basin, lagoon,oxidation ditch, extended aeration, conventional activated sludge,membrane bioreactor, moving bed biofilm reactor, integrated fixed filmactivated sludge, trickle bed reactor, sequencing batch reactor,complete mix, step feed, modified aeration, contact stabilization, highpurity oxygen reactor, Kraus process, or any other reactor for microbialgrowth. In some embodiments, the reaction vessel is an aeration basin.In some embodiments, a wastewater treatment facility comprises theassemblage.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIG. 1 illustrates microorganisms loaded onto an inorganic porousmedium.

FIG. 2 illustrates an example wastewater treatment plant with markedlocations for microorganism delivery and sample extraction.

FIG. 3 illustrates the amount of bleach used at a test facility during atest period and a control period.

FIG. 4 illustrates the Specific Oxygen Uptake Rate (SOUR) at a testfacility during a test period and a control period

FIG. 5 illustrates a percentage of phosphate removed from raw sewage toa secondary treatment effluent.

FIG. 6 illustrates a percentage of phosphate removed from raw sewage toa tertiary treatment in wastewater ponds.

FIG. 7 illustrates total mass of gas produced by microorganisms loadedonto zeolite and microorganisms in a liquid culture.

FIG. 8 illustrates acid produced by microorganisms loaded onto zeoliteand microorganisms in a liquid culture.

FIG. 9 illustrates sugar uptake by microorganisms loaded onto zeoliteand microorganisms in a liquid culture.

DETAILED DESCRIPTION OF THE INVENTION

Reaction vessels are utilized in a wide range of industrial processes,including but not limited to generation of biofuels, treatment of water,food preparation and processing, and manufacturing of biologicalproducts. Industrial reaction vessels are operated in a batch mode, acontinuous process mode, or as a hybrid of batch and continuous processmode. For example, in the manufacture of therapeutic biologicalproteins, batch processing is utilized to obtain stable clinicalproducts at high titers. Continuous bioprocessing is utilized forprocesses that require, for example, an ongoing evolution of a mixedpopulation of cells that are capable of consuming large amounts ofvariable feedstock all year around. Continuous bioprocessing is alsoused in instances where there is production of products that negativelyaffect cell growth or that are unstable and degrade under batchprocessing conditions.

Water, land, and energy resource management continue to be pressingchallenges. Consequently, process optimization of batch, continuous, andhybrid bioprocessing modes is critical to conserving resources andderiving maximum value from current processes utilizing globalresources. Process optimization includes increasing operationalefficiency, increasing carrying capacity of reaction vessel systems, andmaximizing yields while minimizing consumption of raw materials andcosts.

For example, water management involves collection, treatment, andrecycling of both clean water and wastewater. Wastewater treatmentincludes a range of processes such as simple accumulation of wastewaterfollowed by discharge of untreated but screened wastewater streamsdirectly into bodies of water and wastewater treatment plants (WWTP)utilizing sophisticated treatment reaction vessels. The products of thetreatment processes are primarily clean effluent and solids in the formof sludge. Biological treatment of wastewater is accomplished by growingmicrobial species in a continuous reactor mode under aerobic conditions.Wastewater treatment models focus on global growth rates without regardfor the relative abundance of individual species present within thereaction vessel because it is difficult to isolate and accuratelycatalog all species present. Due to the abundance of microbial speciespresent during wastewater treatment, the wastewater treatment industryprovides the most commonly encountered example of complex mixedmicrobial culture interactions.

Process optimizations for wastewater treatment processes include, butare not limited to, reducing the total amount of resulting or producedsludge from the wastewater system that requires disposal orpost-treatment after dewatering, increasing the quality of the resultantbiosolids, and increasing the effluent quality. Technologies used toreduce sludge in wastewater treatment systems include treating withproducts containing enzymatic blends, liquid based microbial cultures,or nutrient based microbial cultures. However, these technologies havebeen unsuccessful in reducing sludge, which consists mostly of water(typically 70% to 85%). Sludge disposal equates to hauling vastquantities of water around the planet every day and utilizes landresources such as energy and fuel. In addition, sludge reduction amountsto significant water conservation as the water content can be returnedto the groundwater supply rather than evaporated, contributing toimpending water shortages.

Key performance indicators that are commonly used in the WWTP industryinclude: amount of mixed liquor suspended solids (MLSS), waste activatedsludge (WAS), volatile suspended solids (VSS), total suspended solids(TSS), recycle ratio, return activated sludge (RAS), biological oxygendemand (BOD), dissolved oxygen (DO), and sludge blanket height. Keyperformance indicators in WWTP focus on suspended and settled solids.One such indicator is the solids retention time (SRT). If the SRT is tooshort, active microorganisms or microorganisms in the log phase may bewashed out. If the SRT is too long, multicellular organisms or undesiredmicroorganism become entrenched and adversely affect the system. Keyperformance indicators from systems that employ various wastewatertreatment systems are shown in Table 1. The values shown in the tableare from a WWTP operator manual (Division of Compliance Assistance.Wastewater Treatment Plant Operator Certification Manual. Frankfort, Ky.Department of Environmental Protection, 2012. Accessed 23 Feb. 2017).

TABLE 1 Design Parameters for Activated Sludge Processes F/M lbs.Organic Return BOD/lb. Loading Detention Flow to Process SRT MLVSS/(lbs. BOD/ MLSS Time Plant Flow Modification (DAYS) day 1000 ft³) (mg/L)(hours)

Conventional 5-15 0.2-0.5 20-40 1000-3000 4-8 0.25-0.75 Complete Mix1-15 0.2-1.0  50-120 1000-6500 3-5 0.25-1.0  Step Feed 3-15 0.2-0.540-60 1500-3500 3-5 0.25-0.75 Modified 0.2-0.5  1.5-5.0  75-150 200-1000 1.5-3.0 0.05-0.25 Aeration Contact 5-15 0.2-0.6 60-751000-3000 0.5-1.0 0.5-1.5 Stabilization 4000-9000 3-6 Extended 20-30 0.05-0.15 12.5-15  2000-6000 18-36 0.5-1.5 Oxidation 10-30  0.05-0.1512.5-15  2000-6000 18-36 0.75-1.5  Ditch High Purity 3-10 0.25-1.0 100-200 3000-8000 1-3 0.25-0.5  Oxygen Kraus Process 5-15 0.3-0.8 40-100 2000-3000 4-8 0.5-1.0

indicates data missing or illegible when filed

Most WWTPs are designed to have a recycle ratio between 50 and 150% ofthe influent flow rate. The typical range for dissolved oxygen, which isthe amount of oxygen that is present in the water, measured inmilligrams per liter, and is usually between 2 and 3.5 mg/L in theaeration basin. In most systems, control is achieved by keeping aconstant MLSS or a constant solids retention time. The MLSS typicallyranges between 2500 and 3500 mg/L. Solids retention time usually rangesbetween 10 and 20 days. The operator alters the wasting rate, which is afraction of the clarifier underflow to keep a steady-state population,measured as MLSS in the basins. The operator maintains a constant sludgeblanket in the clarifiers by changing the RAS or recycle ratio raisingthe RAS flow rate as blanket height climbs and lowering RAS flow rate ifblankets begin to fall.

Most of the VSS into the WWTP (80-90%) are organic foodstuffs likecarbohydrates, lipids, and proteins. A small fraction of the VSS cominginto the WWTP is comprised of non-biodegradable VSS (nbVSS).Approximately 10% of the TSS into the WWTP are comprised of inorganicmaterial like metals and silt. Neither the nbVSS nor the inertinorganics are consumed by biological activity. These solids are not thetarget of activated sludge treatment. The non-biodegradable solids passthrough the WWTP with the majority exiting in the generated sludge and asmall amount remaining suspended and exiting at the outfall per limitsset by the Environmental Protection Agency (EPA). Some fraction, f_(d),of VSS generated in the WWTP remains as non-biodegradable “cell debris.”This cell debris is the major portion of the nbVSS, which along with theinert inorganics comprises sludge and exits the WWTP.

The fraction of total organic carbon (TOC) in VSS that is completelybiodegradable (1-f_(d)) leaves as carbon dioxide, where f_(d) is thenon-biodegradable fraction. The process of waste stabilization involvesthe oxidation of organic material by bacteria with the production ofcarbon dioxide and water. Thus, about 50% of the inbound BOD isconverted to gas (CO₂ and N₂) and water according to equation below.This is called “burn” or “mass to gas.” Consequently, the biomasssynthesis yield is typically less than unity.

${{biomass}{synthesis}{yield}},{Y = \frac{g{biomass}{produced}}{g{substrate}{consumed}}}$

In some embodiments, the yield is defined as:

$Y = \frac{{tons}{of}{dry}{solids}{out}}{{tons}{of}{BOD}{in}}$

Yield varies greatly, but most efficient WWTPs produce around half a tonof sludge for every ton of biodegradable material they receive. Observedyield can be much greater and, in some cases, approach or exceed unity.

Incoming biodegradable material and RAS become nutrients for themicroorganisms in the WWTP. The bacteria either use the nutrients forgrowth (replication) or for cellular maintenance. A small populationprimarily in stationary phase will use the nutrients to maintaincellular functions (catabolism). A large population primarily in loggrowth will use the nutrients to produce more cell mass (anabolism).Given a limited nutrient supply, a larger population will undergo moreendogenous decay (predation on one another) and the decay rate per unittime is increased.

The BOD required to convert organic matter to cell biomass can beapproximated by modeling organic matter as the protein casein. Thechemical formula for casein is C₈H₁₂O₃N₂. Conversion of BOD and organicmatter to cell biomass is represented by the following balanced chemicalequation (Reaction 1):

C₈H₁₂O₃N₂+3O₂→C₅H₇O₂N+3CO₂+H₂O+NH₃

Every 184 grams of organic matter treated will produce 113 grams ofbiomass. This exerts a stoichiometric oxygen demand corresponding tothree moles of oxygen for every mole of organic matter treated. Thisreaction produces approximately 0.61 grams of biomass per gram organicmatter treated. Conversely, 1.42 grams of organic matter is consumed forevery 1 gram of biomass produced. Microbial growth produces off gassingof CO₂ and N₂ and generates water.

The complete oxidation of biomass to carbon dioxide, water, and ammoniais accurately represented by a second balanced chemical equation(Reaction 2):

C₅H₇O₂N+5O₂→5CO₂+2H₂O+NH₃

The first reaction goes essentially to completion (i.e. 100% of theinbound BOD is stabilized and converted to biomass during cell growth).However, the second reaction occurs to the extent that the biodegradablefraction (1-f_(d)) of VSS produced in Reaction 1 is consumed. It is thissecond reaction that converts VSS mass to gas, thereby reducing theobserved yield of outbound solids further below the 60% biosolids yieldfrom equation above. The mass fraction of TOC in BOD is 96/184 or 52%,signifying that carbon makes up over 50% of the total BOD mass to betreated. Similarly, there is available oxygen contained in BOD. The massfraction of BOD that is oxygen is 48/184 or 26%.

Carbon does not accumulate, but leaves the WWTP as either gas or sludge.The fraction of TOC in BOD that leaves as gas is 36/96 or 37.5%. Thebalance of the TOC in BOD that remains captured in biomass is 60/96 or62.5%. It is the remaining biomass that can be further reduced to gas.Of the mass of BOD into the reactor, almost 10% is reduced to liquidwater in the basins. A significant amount of water is generated duringmicrobial growth. Organic nitrogen comprises 28/184 or 15% of the BODmass load. Half of the organic nitrogen is converted throughnitrification/denitrification to molecular nitrogen in Reaction 1.Including the TOC and nitrogen that leave the WWTP as gas, approximately45% of the total inbound organic load to the WWTP is lost as gas as aresult of the first reaction alone. The about 45% reduction of BOD massinbound indicates that the stabilization process (Reaction 1) aloneachieves a biomass synthesis yield of around 55%. Improvement in thereduction of the yield from the process occurs as the resulting biomassis further oxidized and gasified through Reaction 2. Reaction 2describes endogenous decay of the biomass generated within the WWTP.Reaction 2, if complete, converts all the biomass generated into gas,water, and ammonia. In the case of complete conversion, sludge wouldcontain only inert inorganics and inert VSS that had entered the WWTP inthe influent. Influent streams with greatly different compositionalcharacterization, such as higher loading concentrations of BOD andchemical oxygen demand (COD) and streams with much higher inert fractionin the influent may show much higher biomass synthesis yields than the50% to 60% range described here.

Disclosed herein, in certain embodiments, are compositions, methods, andassemblages for enhancing effluent and biosolids quality for reactionvessels and wastewater treatment facilities using a microbial species,or a consortium of microbial species, loaded onto a porous medium.

Certain Definitions

As used herein, “reaction vessel” refers to any system containingmicroorganisms, in which materials are converted by the microorganisms,products produced by the microorganisms, or in which increase cellpopulation is achieved. Reaction vessels used herein can be one or moreof batch reactors, fed-batch reactors, semi-continuous reactors,continuous stirred-tank reactors, continuous flow stirred-tank reactors,and plug-flow reactors, singularly or in series; ebullized-bed (i.e.,“bubbling and boiling”) reactors; and fluidized-bed reactors. In certainembodiments, the reaction vessel can be an aeration basin or lagoon. Insome embodiments, the reaction vessel can be one or more of a tricklingbed reactor, percolating reactors, fluidized reactor, plug-flowreaction, counter-current reactor, sequencing batch reactor, rotatingbiological contactors, oxidation ditch, extended aeration, conventionalactivated sludge, membrane bioreactor, moving bed biofilm reactor,integrated fixed film activated sludge, trickle bed reactor, sequencingbatch reactor, complete mix, step feed, modified aeration, contactstabilization, high purity oxygen reactor, or Kraus process.

As used herein, “wastewater treatment” refers to a process that convertswater that is contaminated water or unsuitable for consumption by plantsor animals into effluent and biosolids that can be reused for anotherpurpose or returned to the water cycle.

As used herein, the phrase “inorganic porous medium” refers to aninorganic support having a porous structure. In some embodiments, theinorganic porous medium is precipitated silica granules, super absorbentsilica polymers, crystalline silica, fused quartz, fumed silica, silicagels, aerogels, colloidal silica, zeolite, aluminosilicate, silicate,diatomaceous earth, or alumina. In some embodiments, zeolite comprisesandalusite, kyanite, sillimanite, analcime, chabazite, clinoptilite,mordenite, natrolite, heulandite, phillipsite, or stilbite. In someembodiments, the inorganic porous medium is a mixture of different typesof inorganic porous mediums. In some embodiments, the porous structureis loaded with least one microbial species.

As used herein, “delivered microorganisms” refers to bacteria, viruses,mycoplasma, fungi, and protozoa loaded onto an inorganic porous medium.In some embodiments, the microorganisms loaded onto the inorganic porousmedium are bacteria. In some embodiments, the microorganisms include asingle species of microorganism or a consortium of microorganisms. Insome embodiments, the microorganism(s) are selected based on intendeduse, the available nutrient sources, and the desired operatingconditions of the reaction vessel.

As used herein, “carrying capacity” refers to the maximum populationthat a particular reaction vessel system can support. In a continuousreactor system, such as a WWTP, the carrying capacity is measured astotal suspended solids (TSS), mixed liquor suspended solids (MLSS), orvolatile suspended solids (VSS). In some examples, the carrying capacityis measured by the increased rate of consumption of glucose or othersugars. In a batch reactor system, the carrying capacity is measured bythe peak population density of microorganisms or by measuring the rateof growth of the microorganisms and the rate of nutrient consumption.

Delivered Microorganisms

Disclosed herein, in certain embodiments, are methods of deliveringmicroorganisms. FIG. 1 shows an inorganic porous medium with suitablecharacteristics for microorganism loading. In some embodiments,characteristics for microorganism loading includes, but is not limitedto, loading capability, mineral content, pore size, chemical inertness,and porosity. In some embodiments, a microbial solution containing asingle microbial species and nutrients necessary for growth of themicrobial species is loaded onto an inorganic porous medium. In someembodiments, a microbial solution containing a consortium of microbialspecies and nutrients necessary for growth of the microbial species isloaded onto an inorganic porous medium. In some embodiments, theaddition of the microbial solution to the inorganic porous mediumproduces a substance that is dry-to-the-touch creating a dry mode fordelivery of microorganism. In some embodiments, the inorganic porousmedium comprises zeolite. In some embodiments, the zeolite comprises,but is not limited to, andalusite, kyanite, sillimanite, analcime,chabazite, clinoptilite, mordenite, natrolite, heulandite, phillipsite,or stilbite. In some embodiments, the inorganic porous medium comprisesaluminosilicate, silicate, or diatomaceous earth. In some embodiments,the inorganic porous medium is a mixture of different types of inorganicporous mediums. In some embodiments, the inorganic porous mediumcomprises precipitated silica granules. In some embodiments,precipitated silica granules are highly porous and contain a largesurface area both within their volume and on the surface. In someembodiments, one pound of silica granules has approximately 700,000square feet of surface area. In some embodiments, the surface areaprovides a matrix upon which a reaction can be accelerated. In someembodiments, precipitated silica granules are also a super absorbentpolymer capable of drawing in organic nutrients to be used as buildingblocks for new bacterial cells and to sustain cellular functions. Insome embodiments, as the microorganisms reach exponential growth phasewithin an inorganic porous medium, they experience crowding effectswithin the medium and populate the surrounding environment.

In some embodiments, the microorganism comprises a mixed culture ofbeneficial microbes. In some embodiments, the microorganisms comprise anative, non-pathogenic consortium of microbial species ideal forwastewater applications. In some embodiments, the microbial species arenot genetically modified strains. In some embodiments, the microbialspecies belong to the class of Group 1 microorganisms according to theWorld Health Organism (WHO), where Group 1 microorganisms aremicroorganisms unlikely to cause disease. In some embodiments,microorganisms loaded onto an inorganic porous medium produce blooms ofbeneficial bacteria when placed into an aqueous environment containing anutrient source in the form of biomass or dead cells.

In some embodiments, the microorganisms loaded onto an inorganic porousmedium are delivered to a reaction vessel. In some embodiments, themicroorganisms, the nutrients required for optimal growth, and theinorganic porous medium are delivered independently to a reactionvessel. In some embodiments, the microorganisms loaded onto an inorganicporous medium are delivered to a batch reactor. In some embodiments, themicroorganisms loaded onto an inorganic porous medium are delivered to acontinuous reactor. In some embodiments, the microorganisms loaded ontoan inorganic porous medium are delivered to a hybrid of a batch reactorand a continuous reactor. In some embodiments, the reaction vessels arecarried out under aerobic or anaerobic conditions depending on thereaction and microorganism(s) involved. In some embodiments,microorganisms loaded onto an inorganic porous medium are used toproduce biofuels, including but not limited to methanol, ethanol, orbutanol. In some embodiments, microorganisms loaded onto an inorganicporous medium are used to produce biogas. In some embodiments,microorganisms loaded onto an inorganic porous medium are used toenhance wastewater treatment. In some embodiments, microorganisms loadedonto an inorganic porous medium are used to produce amino acids. In someembodiments, microorganisms loaded onto an inorganic porous medium areused to produce therapeutically important peptides.

Microorganism Delivery to a WWTP

Disclosed herein, in certain embodiments, are methods of producing andcompositions of a fertilizer or compost derived from a reaction vessel.In some embodiments, microorganisms loaded onto an inorganic porousmedium are delivered to a reaction vessel. In some embodiments, themicroorganisms are aerobic, anaerobic, or facultative. In someembodiments, the reaction vessel comprises an influent stream, aneffluent stream, an aqueous phase, and a biosolids phase. In someembodiments, the biosolids phase comprises nutrients for themicroorganisms. In some embodiments, the effluent stream is separatedinto an aqueous phase and a biosolids phase. In some embodiments, thebiosolids phase is dewatered to produce a fertilizer or compost. In someembodiments, the biosolids phase is digested in a digester prior todewatering. In some embodiments, the influent stream includes, but isnot limited to, residential wastewater, agricultural wastewater,industrial wastewater, runoff wastewater or any combination thereof. Insome embodiments, adding microorganisms loaded onto an inorganic porousmedium to a reaction vessel increases the carrying capacity of thereaction vessel. In some embodiments, adding microorganisms loaded ontoan inorganic porous medium to a reaction vessel reduces the quantity ofthe biosolids phase. In some embodiments, separating the effluent streaminto the aqueous phase and the biosolids phase includes, but is notlimited to, decantation, filtration, centrifugation, or any combinationthereof. In some embodiments, dewatering the biosolids phase includescentrifugation and filtration.

In some embodiments, the reaction vessel is part of a WWTP. In someembodiments, the reaction vessel is an aeration basin, lagoon, or lake.In some embodiments, the WWTP is modeled as a chemostat. In someembodiments, the microorganisms are bacterial. In some embodiments, theWWTP must produce as many bacteria as wash out in the effluent stream.In some embodiments, washing out of the bacteria leads to a WWTP devoidof beneficial bacteria. In some embodiments, growth rate of a singlemicrobial species within a WWTP are described by Michaelis-Mentenkinetics. In some embodiments, the bacteria will grow exponentiallyuntil the food source is depleted and crowding occurs. In someembodiments, exponential growth is log-linear and corresponds to a shortdoubling time for the microbial population. In some embodiments,nutrient consumption is rapid during exponential growth. In someembodiments, when the reaction vessel carrying capacity is reached, themicroorganisms enter a stationary phase. In some embodiments, during thestationary phase the number of microorganisms produced is equal to thenumber of microorganisms consumed and the overall population remainsunchanged. In some embodiments, during the stationary phase substrateuptake corresponds to a “maintenance” requirement. In some embodiments,the microbial population in a WWTP consists primarily of microorganismin the stationary phase. In some embodiments, the nutrients have beendepleted and the microorganism population begins to decline byendogenous decay. In some embodiments, endogenous decay involves celllysis and the conversion of dead cell mass into nutrients for otherviable microorganisms. In some embodiments, the activated sludge isrecycled back to the aeration basin to allow the dead cells to becomenutrients for younger microorganisms. In some embodiments,microorganisms loaded onto an inorganic porous medium drive the WWTPtowards greater endogenous decay and cause more mass to leave as gas. Insome embodiments, higher microbial activity means more highly treatedwater.

In some embodiments, the inorganic porous medium is selected or modifiedfor sorption of nitrogen, phosphorous, or both nitrogen or phosphorousfrom an aqueous phase of a reaction vessel. In some embodiments, thephosphorous is an organic or an inorganic aqueous phosphorous. In someembodiments, sorption of nitrogen and/or phosphorous from an aqueousphase of a reaction vessel increases a nutrient concentration of afertilizer or compost produced from the reaction vessel. In someembodiments, sorption of nitrogen and/or phosphorous from an aqueousphase of a reaction vessel decreases the amount of nitrogen and/orphosphorous in the aqueous phases. In some embodiments, the inorganicporous medium may be chemically or physically modified. In someembodiments, chemical modification includes the addition of chelatingagents, ligands, or salts (e.g., magnesium salts) that lead toprecipitation of compounds that referentially bind ammonia andphosphorous. In some embodiments, physical modification includesroughening the surfaces of the porous medium, increasing porosity of theporous medium, inducing the formation of aggregates or agglomerates fromsmaller particles of the inorganic porous media using coagulation andflocculation agents, or any combination thereof. In some embodiments, aflocculating agent is added to the effluent stream to produce an aqueousphase and a filter cake. In some embodiments, the flocculating agent isone or more of an ionic polymer, a non-ionic polymer, or any combinationthereof. In some embodiments, the ionic polymer is a cationic polymer.In some embodiments, the ionic polymer is an anionic polymer. In someembodiments, flocculating agents include aluminum chloride, ferricchloride, and alum. In some embodiments, the cationic polymer is acopolymer of AETAC (N,N-Dimethylaminoethyl Acrylate Methyl ChlorideQuaternary) or METAC (N,N-Dimethylaminoethyl Methacrylate MethylChloride Quaternary) and acrylamide. In some embodiments, flocculatingagents perform a dual function of coagulating with their positive ioniccharge and flocculating with their high molecular weight. In someembodiments, the anionic polymer is a copolymer of acrylamide andacrylic acid. In some embodiments, there is at least about a 45%reduction in the consumption of flocculating agents, when compared tosystems that do not employ delivered microorganisms. In someembodiments, there is at least about a 40% reduction in the consumptionof flocculating agents. In some embodiments, there is at least about a35% reduction in the consumption of flocculating agents. In someembodiments, there is at least about a 30% reduction in the consumptionof flocculating agents. In some embodiments, there is at least about a25% reduction in the consumption of flocculating agents. In someembodiments, there is at least about a 20% reduction in the consumptionof flocculating agents.

In some embodiments, the concentration of MLSS in the effluent isgreater than about 7,000 mg/L. In some embodiments, the concentration ofMLSS in the effluent is greater than about 8,000 mg/L. In someembodiments, the concentration of MLSS in the effluent is greater thanabout 9,000 mg/L. In some embodiments, the concentration of MLSS in theeffluent is greater than about 10,000 mg/L. In some embodiments, theconcentration of MLSS in the effluent is greater than about 11,000 mg/L.In some embodiments, the concentration of MLSS in the effluent isgreater than about 12,000 mg/L. In some embodiments, increasing the MLSSis an important measure for determining the load delivered to a solidseparator, such as a clarifier. In some embodiments, the settlingcharacteristics of the MLSS vary from system to system. In someembodiments, the settling characteristics of the MLSS determine theupper boundary of solids concentration of MLSS being fed to a clarifieror other type of solid liquid separator. In some embodiments, the solidliquid separator surface area and the mass rate of suspended solidsintroduced to the clarifier are used to determine the mass flux. In someembodiments, the mass flux is a process design parameter for determiningthe operational size of the clarifier. In some embodiments, a higherMLSS also has a higher VSS. In some embodiments, a higher VSS indicateshigher beneficial microbial activity in the WWTP operations.

In some embodiments, the SRT is greater than twenty days. In someembodiments, the SRT is greater than thirty days. In some embodiments,the SRT is greater than forty days. In some embodiments, the SRT isgreater than forty-five days. In some embodiments, the SRT of the sludgeor solids is greater than fifty days. In some embodiments, the SRT isgreater than sixty days. In some embodiments, the SRT is increased bygreater than or equal to about 1%, 2%, 5%, 10%, 20%, 25%, 30%, 40%, 50%,60%, 70%, 80%, 90%, 100%, 150%, 200%, 250%, 300%, or more when thedelivered microorganism is used in a WWTP.

In some embodiments, the SRT of the solids is maintained constantbetween a WWTP that does not use delivered microorganisms and a systemthat does use delivered microorganisms. In some embodiments, when SRT ofthe solids is maintained constant, there is at greater than or equal toa 2.5% reduction in solids or sludge produced, as compared to the samesystem without the use of delivered microorganisms. In some embodiments,when SRT of the solids is maintained constant, there is greater than orequal to a 5% reduction in solids or sludge produced, as compared to thesame system without the use of delivered microorganisms. In someembodiments, when SRT of the solids is maintained constant, there isgreater than or equal to a 7.5% reduction in solids or sludge produced,as compared to the same system without the use of deliveredmicroorganisms. In some embodiments, when SRT of the solids ismaintained constant, there is greater than or equal to a 10% reductionin solids or sludge produced, as compared to the same system without theuse of delivered microorganisms. In some embodiments, when SRT of thesolids is maintained constant, there is greater than or equal to a 15%reduction in solids or sludge produced, as compared to the same systemwithout the use of delivered microorganisms. In some embodiments, whenSRT of the solids is maintained constant, there is greater than or equalto a 20% reduction in solids or sludge produced, as compared to the samesystem without the use of delivered microorganisms. In some embodiments,when SRT of the solids is maintained constant, there is greater than orequal to a 25% reduction in solids or sludge produced, as compared tothe same system without the use of delivered microorganisms. In someembodiments, when SRT is maintained constant, there is greater than orequal to a 30% reduction in solids or sludge produced, as compared tothe same system without the use of delivered microorganisms. In someembodiments, when SRT of the solids is maintained constant, there isgreater than or equal to a 40% reduction in solids or sludge produced,as compared to the same system without the use of deliveredmicroorganisms.

In some embodiments, the sludge produced at a WWTP is reduced by atleast about 40% as compared to systems that do not employ deliveredmicroorganisms. In some embodiments, the sludge produced at a WWTP isreduced by at least about 30%. In some embodiments, the sludge producedat a WWTP is reduced by at least about 25%. In some embodiments, thesludge produced at a WWTP is reduced by at least about 20%. In someembodiments, the sludge produced at a WWTP is reduced by at least about15%. In some embodiments, the economic benefit of sludge reductionincludes financial savings, time savings, a reduction in personnelresources associated with sludge disposal, reduction in consumption offlocculating agents such as polymers, increase in equipment life, andreduction in equipment maintenance costs. In some embodiments, areduction in WWTP operational costs includes lower oxygen demand, higherblower efficiency, decreased qualitative and quantitative use ofchemicals for sanitation, and extended WWTP life. Key operatingparameters for a WWTP system employing delivered microorganisms is shownin Table 2.

TABLE 2 Design Parameters for Activated Sludge Processes F/M lbs.Organic BOD/lb. Loading Detention Return Process SRT MLVSS/ (lbs. BOD/MLSS Time Flow to Modification (DAYS) day 1000 ft³) (mg/L) (Hours) PlantFlow Delivery 30-50 0.5-0.05 20-200 7,000-11,000 4-17 .5-2.0microorganisms

In some embodiments, the yield which is measured as a unit mass of wasteproduced per unit mass of organic loading is less than about 40%. Insome embodiments, the yield is less than about 30%. In some embodiments,the yield is less than about 20%. In some embodiments, lower yieldrepresents minimization or reduction of biosolids for wastewatertreatment.

In some embodiments, the dewatered biosolids phase produced after theaddition of microorganisms loaded onto an inorganic porous medium is ofan enhanced quality compared to a biosolids phase produced without theaddition of microorganisms loaded onto an inorganic porous medium. Insome embodiments, the enhanced dewatered biosolids contain at least1,500 ppm, 1,250 ppm, 1,000 ppm, 750 ppm, 500 ppm, 250 ppm, or fewer ppmof an inorganic porous medium on a dry basis. In some embodiments, theenhanced dewatered biosolids contain at least about 500 ppm of aninorganic porous medium on a dry basis. In some embodiments, theenhanced dewatered biosolids contain less than about 150 ppm, 125 ppm,100 ppm, 75 ppm, 50 ppm, 25 ppm, or fewer ppm arsenic. In someembodiments, the enhanced dewatered biosolids contain less than about 41ppm arsenic. In some embodiments, the enhanced dewatered biosolidscontain less than about 150 ppm, 125 ppm, 100 ppm, 75 ppm, 50 ppm, 25ppm, or fewer ppm cadmium. In some embodiments, the enhanced dewateredbiosolids contain less than about 39 ppm cadmium. In some embodiments,the enhanced dewatered biosolids contain less than about 2,500 ppm,2,000 ppm, 1,500 ppm, 1,000 ppm, 500 ppm, or fewer ppm chromium. In someembodiments, the enhanced dewatered biosolids contain less than about1,200 ppm chromium. In some embodiments, the enhanced dewateredbiosolids contain less than about 3,000 ppm, 2,500 ppm, 2,000 ppm, 1,500ppm, 1,000 ppm, 500 ppm, or fewer ppm copper. In some embodiments, theenhanced dewatered biosolids contain less than about 1,500 ppm copper.In some embodiments, the enhanced dewatered biosolids contain less thanabout 1000 ppm, 750 ppm, 500 ppm, 250 ppm, or fewer ppm lead. In someembodiments, the enhanced dewatered biosolids contain less than about300 ppm lead. In some embodiments, the enhanced dewatered biosolidscontain less than about 100 ppm, 75 ppm, 500 ppm, 25 ppm, 15 ppm, orfewer ppm mercury. In some embodiments, the enhanced dewatered biosolidscontain less than about 17 ppm mercury. In some embodiments, theenhanced dewatered biosolids contain less than about 1000 ppm, 500 ppm,400 ppm, 300 ppm, 200 ppm, or fewer ppm nickel. In some embodiments, theenhanced dewatered biosolids contain less than about 420 ppm nickel. Insome embodiments, the enhanced dewatered biosolids contain less thanabout 150 ppm, 100 ppm, 75 ppm, 50 ppm, 25 ppm, or fewer ppm selenium.In some embodiments, the enhanced dewatered biosolids contain less thanabout 36 ppm selenium. In some embodiments, the enhanced dewateredbiosolids contain less than about 7,500 ppm, 5,000 ppm, 2,500 ppm, 1,000ppm, or fewer ppm zinc. In some embodiments, the enhanced dewateredbiosolids contain less than about 2,800 ppm zinc.

In some embodiments, the enhanced dewatered biosolids contains aconcentration of Salmonella enterica spp. less than about 10 mostprobable number (MPN), 8 MNP, 5 MPN, 3 MPN, or fewer MPN. In someembodiments, the enhanced dewatered biosolids contain less than about 3MPN Salmonella enterica spp. In some embodiments, the enhanced dewateredbiosolids contains a total concentration of fecal coliform bacteria ofless than about 2,000 MPN, 1,500 MPN, 1,250 MPN, 1,000 MPN, 750 MPN, orfewer MPN. In some embodiments, the enhanced dewatered biosolids containless than about 1000 MPN of fecal coliform. In some embodiments, theenhanced dewatered biosolids contain less than about 10 plaque-formingunit (PFU), 8 PFU, 4 PFU, 2 PFU, 1 PFU, or less PFU of enteric virusesper four grams of total solids on a dry basis. In some embodiments, theenhanced dewatered biosolids contain less than about 1 PFU of entericviruses per four grams of total solid on a dry basis. In someembodiments, enteric viruses include human astroviruses, humanadenoviruses, noroviruses, human sapoviruses, human parvoviruses,non-polio enteroviruses, and human rotoviruses. In some embodiments, theenhanced dewatered biosolids contain less than about 10, 8, 4, 2, 1 orless viable helminth ova per four grams of total solids on a dry basisand with a vector attraction standard oxygen uptake rate of less thanabout 5 mg, 4 mg, 3 mg, 1.5 mg, or less mg of oxygen per gram of solidsper hour. In some embodiments, the enhanced dewatered biosolids containless than about 1 viable helminth ova per four grams of total solids ona dry basis and with a vector attraction standard oxygen uptake rate ofless than about 1.5 mg of oxygen per gram of solids per hour.

In some embodiments, the enhanced dewatered biosolids comprise less thanabout 41 ppm arsenic, less than about 39 ppm cadmium, less than about1,200 ppm chromium, less than about 1,500 ppm copper, less than about300 ppm lead, less than about 17 ppm mercury, less than about 420 ppmnickel, less than about 36 ppm selenium, and less than about 2,800 ppmzinc; a concentration of Salmonella enterica spp. below about 3 mostprobable number (MPN) per 4 grams of total solids on a dry basis; atotal concentration of fecal coliform bacteria less than about 1000 MPNper gram of total solids on a dry basis; a density of enteric virusesless than about 1 plaque-forming unit (PFU) per 4 grams of total solidson a dry basis; and/or a density of viable helminth ova less than about1 per 4 grams of total solids on a dry basis with a vector attractionstandard oxygen uptake rate of less than about 1.5 milligrams of oxygenper gram of solids per hour.

In some embodiments, the enhanced dewatered biosolids comprise no morethan 41 ppm arsenic, no more than 39 ppm cadmium, no more than 1,200 ppmchromium, no more than 1,500 ppm copper, no more than 300 ppm lead, nomore than 17 ppm mercury, no more than 420 ppm nickel, no more than 36ppm selenium, and no more than 2,800 ppm zinc; a concentration ofSalmonella enterica spp. no more than 3 most probable number (MPN) per 4grams of total solids on a dry basis; a total concentration of fecalcoliform bacteria no more than 1000 MPN per gram of total solids on adry basis; a density of enteric viruses no more than 1 plaque-formingunit (PFU) per 4 grams of total solids on a dry basis; and/or a densityof viable helminth ova no more than 1 per 4 grams of total solids on adry basis with a vector attraction standard oxygen uptake rate of nomore than 1.5 milligrams of oxygen per gram of solids per hour.

In some embodiments, the effluent stream produced after the addition ofdelivered microorganisms is of an enhanced quality compared to aneffluent produced without the addition of microorganisms loaded onto aninorganic porous medium. In some embodiments, the effluent stream has areduced ammonia concentration. In some embodiments, the ammoniaconcentration is less than about 1 mg/L, 0.75 mg/L, 0.5 mg/L, 0.25 mg/L,0.1 mg/L, or fewer mg/L. In some embodiments, the ammonia concentrationis less than about 0.2 mg/L. In some embodiments, the ammoniaconcentration is less than the analytical detection limit. In someembodiments, the reaction vessel is operated under aerobic conditions.In some embodiments, the microorganisms consume ammonia under aerobicconditions to produce nitrite and nitrate. In some embodiments, theammonia is converted to nitrite and nitrate. In some embodiments, theconcentration of nitrates and nitrites is less than about 50 mg/L, 40mg/L, 30 mg/L, 20 mg/L, 10 mg/L, 5 mg/L, 2.5 mg/L, or fewer mg/L. Insome embodiments, the concentration of nitrates and nitrites is in therange of about 5 mg/L to 30 mg/L. In some embodiments, the effluentstream is denitrified. In some embodiments, the reaction vessel isoperated under anaerobic conditions. In some embodiments, themicroorganisms consume nitrate and nitrate under anaerobic conditions toproduce molecular nitrogen. In some embodiments, a reduction in ammoniaand denitrification does not require a chlorinator, ozonator, or the useof a peroxide, bleach or ultra-violet light. In some embodiments, thephosphorous concentration in the effluent is reduced. In someembodiments, the concentration of phosphorous in the effluent is lessthan about 20 mg/L, 10 mg/L, 5 mg/L, 2.5 mg/L, or fewer mg/L. In someembodiments, the concentration of phosphorus in the effluent is lessthan about 3 mg/L. In some embodiments, the microorganisms consumephosphorous. In some embodiments, the consumed phosphorous isincorporated into cell biomass. In some embodiments, the removal ofsoluble phosphorous reduces or prevents eutrophication. In someembodiments, phosphorous removal does not require phosphateprecipitation with calcium, aluminum, iron, or magnesium. In someembodiments, phosphorous removal is achieved under anaerobic conditions.In some embodiments, phosphorous removal is achieved under aerobicconditions.

WWTP Using Delivered Microorganisms

Disclosed herein, in certain embodiments, are assemblages for treatingwastewater using delivered microorganisms. FIG. 2 shows an example WWTPwith example locations to add delivered microorganisms and locations toextract test samples. In some embodiments, the delivered microorganismsare added to the inlet stream before the aeration basin 201 or to theaeration basin 202. In some embodiments, test samples are extracted fromthe influent stream 210. In some embodiments, test samples are extractedfrom the effluent 220. In some embodiments, test samples are extractedfrom the clarifier overflow 230. In some embodiments, test samples areextracted after the digester 240. In some embodiments, test samples areextracted from the filter cake 250. In some embodiments, test samplesare extracted from or after the aeration basin 260. In some embodiments,test samples are analyzed for total volatile suspended solids, ammonia,COD, TSS, BOD, nitrate and nitrite as elemental nitrogen, phosphorous,and alkalinity. In some embodiments, total volatile suspended solids areanalyzed following EPA method 160.4 (Environmental Protection Agency.(1971). Method 160.4: Residue, Volatile (Gravimetric, Ignition at 550°C.) by Muffle Furnace). In some embodiments, ammonia is analyzedfollowing EPA method 350.1 (Environmental Protection Agency (1993).Method 350.1: determination of ammonia nitrogen by Semi-AutomatedColorimetry). In some embodiments, COD is analyzed following an EPAapproved method supplied by the Hach Company, HACH 8000. In someembodiments, TSS are analyzed following Standard Methods 2540D (Eaton,A. D., Clesceri, L. S., Greenberg, A. E., Franson, M. A. H., AmericanPublic Health Association, American Water Works Association, and WaterEnvironment Federation. (2012). Standard methods for the examination ofwater and wastewater. Washington, D.C.: American Public HealthAssociation). In some embodiments, BOD is analyzed following StandardMethods 5210B. In some embodiments, nitrates and nitrates are analyzedas elemental nitrogen following EPA method 300.0 (EnvironmentalProtection Agency (1993). Method 300.0: determination of inorganicanions by ion chromatography). In some embodiments, phosphorous isanalyzed by Standard Method 4500P. In some embodiments, Alkalinity isanalyzed by Standard Methods 2320A.

In some embodiments, an assemblage for treating wastewater comprises oneor more influent streams, one or more aeration basins or aerationlagoons, one or more separators, and one or more effluent streams. Insome embodiments, the influent stream comprises an aqueous phase and abiosolids phase. In some embodiments, the influent stream includesresidential wastewater, agricultural wastewater, industrial wastewater,runoff wastewater, or any combination thereof. In some embodiments, theaeration basins comprise at least one inlet stream and at least oneoutlet stream. In some embodiments, the separator separates the aqueousphase from the biosolids phase. In some embodiments, the deliveredmicroorganisms are added upstream of the aeration basin. In someembodiments, the delivered microorganisms are added to the aerationbasin or aeration lagoon. In some embodiments, the aeration basin isconfigured to mix wastewater with the microorganism loaded onto theinorganic porous medium. In some embodiments, the wastewater treatmentassemblage has an increased carrying capacity. In some embodiments, theoutlet stream of the aeration basin has a microbial activity at leasttwice that of the inlet stream.

In some embodiments, a first separator is configured to receivewastewater from the aeration basin or aeration lagoon to produce a firstfraction containing biosolids and a treated water stream. In someembodiments, a second separator is configured to receive the firstfraction containing biosolids to produce a second fraction containingbiosolids and a waste product stream containing biosolids. In someembodiments, the second fraction is recycled to the aeration basin. Insome embodiments, the assemblage includes a third separator configuredto produce a treated water stream and a filter cake. In someembodiments, a biosolids fraction from the first, second, or thirdseparator is directed to a reaction vessel to produce digested products.In some embodiments, the reaction vessel that produces digested productsin a digester. In some embodiments, the digested products are directedto an additional separator configured to remove the aqueous phase fromthe digested product and produce a filter cake. In some embodiments, aRAS line carries a portion of the biosolids phase from the separatorback to the aeration basin. In some embodiments, the portion of thebiosolids phase returned to the aeration basin increases the TSS in theaeration basin by 25%. In some embodiments, the portion of the biosolidsphase returned to the aeration basin increases the TSS in the aerationbasin by 50%. In some embodiments, the portion of the biosolids phasereturned to the aeration basin increases the TSS in the aeration basinby 100%. In some embodiments, the concentration of MLSS is greater than7000 g/L. In some embodiments, SRT in the aeration basin is greater thantwenty days. In some embodiments, a WAS line caries a second portion ofthe biosolids phase to a dewaterer that dewaters the biosolid phase. Insome embodiments, the assemblage does not comprise a chlorinator or anozonator. In some embodiments, the separator comprises a decanter,filter, centrifuge, or combination thereof.

EXAMPLES Example 1: Biosolids Reclassification Based on Pathogen Limitsand Vector Attraction Based on Standard Oxygen Uptake Rates

A test facility was selected at the Kingwood-Central wastewatertreatment facility in Kingwood, Tex. This specific plant has anexcellent record of meeting compliance. The Kingwood-Central wastewatertreatment facility has an average daily flow rate of 5.0 million gallonsof wastewater. The test began in March of 2015 with the introduction ofa microbial species loaded onto an inorganic porous medium, or deliveredmicroorganisms, to the aeration basins of the plant at a rate of 15pounds per day. The delivered microorganisms were applied either onceper day or applied in three hour increments until a total of 15 poundswas added. An independent contractor collected and performed sampleanalytics that provide data acquisition over the entire test period.These additional samples and analytics augmented the standard analyticsperformed by the operator. The test period covered a total of 21 months,from March 2015 through October 2016.

Improved Filter Cake Quality

The performance of the delivered microorganisms show a reduction offecal coliform to 1,100 or less cfg/g of total solids and a vectorattraction based on Standard Oxygen Uptake Rate (SOUR) ranging from 0.24to 0.39 mg of oxygen per gram of solids per hour.

Analytical results prior to the testing period are shown in Table 3 forthe period starting in October 2013 and ending in December 2014. Datafrom the historical operation of the plant is used as a baseline forcomparison.

TABLE 3 Analytical Results from October 2013 to December 2014. Sampledate Oct. 10, 2013 Sep. 2, 2014 Dec. 23, 2014 Parameters Cake Cake CakepH (Units) 6.57 7.19 7.47 Tot. Nitrogen-N (%) 0.53 0.53 0.64 NO3-N (%)0.02 0.02 0.01 NH3-N (%) 0.01 0.4 0.03 Phosphorus pentoxide (%) 0.021.13 1.05 Potassium % 0.24 0.08 0.13 Arsenic (mg/kg) 4.21 3.7 2.45Cadmium (mg/kg) 1.75 1.23 0.61 Copper (mg/kg) 781.96 608.8 422.49Molybdenum (mg/kg) 14.38 11.11 9.81 Nickel (mg/kg) 16.83 12.75 9.2 Lead(mg/kg) 28.4 25.5 15.94 Chromium (mg/kg) 26.65 20.57 13.49 Mercury(mg/kg) 0.07 0.08 0.12 Selenium (mg/kg) 8.77 5.76 5.52 Zinc (mg/kg)1171.19 1028.38 766.49 PCB's (mg/kg) 1 1 1 Fecal coliform (cfg/g/TS)12,900 110,000 5,700 SOUR (mg O2/g/hr) 0.3 0.25 1.2 Total solids (%)14.7 18.7 15.1 Volatile solids (%) 7.7 9.2 9 Organic concentration (%)52.4 49.2 59.6

Analytical results from the test period are shown in Table 4. Use of amicrobial species loaded onto an inorganic porous medium decreased thefecal coliform.

TABLE 4 Analytical Results from August 2015 to July 2016. Sample dateAug. 15, 2015 Jul. 31, 2016 Parameters Cake Cake pH (Units) 6.75 6.27Tot. Nitrogen-N (%) 0.58 0.18 NO3-N (%) 0.03 0.05 NH3-N (%) 0.05 0.03Phosphorus pentoxide (%) 1.36 0.73 Potassium % 0.18 0.15 Arsenic (mg/kg)2.02 0.64 Cadmium (mg/kg) 2.02 1.93 Copper (mg/kg) 597.1 633.07Molybdenum (mg/kg) 10.76 9.03 Nickel (mg/kg) 12.1 12.89 Lead (mg/kg)26.9 20.63 Chromium (mg/kg) 26.22 20.63 Mercury (mg/kg) 0.13 0.13Selenium (mg/kg) 6.05 7.74 Zinc (mg/kg) 974.99 1205.55 PCB's (mg/kg) 1 1Fecal coliform (cfg/g/TS) <1000 1,100 SOUR (mg O2/g/hr) 0.39 0.24 Totalsolids (%) 14.3 15.7 Volatile solids (%) 7.6 7.52 Organic concentration(%) 53.1 47.9

Ammonia Reduction

Samples were taken at the inlet to the plant, at the clarified overflow,from the digester, and from the aqueous effluent from the plant. Ammoniasamples were analyzed by EPA Method 350.1 and Nitrates were analyzed byEPA method 352.1 (Environmental Protection Agency. (1971). Method 350.1:Nitrogen, Nitrate). Analysis results are shown in Table 5 and Table 6for ammonia and nitrate, respectively. Use of a microbial species loadedonto an inorganic porous medium reduced the ammonia at the clarifieroverflow by 96% and at the digester by 75%.

TABLE 5 Analytical Results of Ammonia Concentration. (in mg/l) AmmoniaInfluent Clarifier OF Digester Effluent Baseline 30 13 13 0.2 Delivered31 0.6 3 0.2 Microorganisms

TABLE 6 Analytical Results of Nitrate Concentration. (in mg/l)Nitrate/Nitrite Influent Clarifier OF Digester Effluent Baseline 0.7 170.6 17 Delivered 0.7 19 1.2 18 Microorganisms

Nitrification and Denitrification in the Digester

The performance of the delivered microorganisms shows a 90% reduction oftotal nitrogen as ammonia and nitrate/nitrite compared to the baselinereduction of 70% when comparing the WAS concentrations to the existingdigester concentrations. The ammonia was reduced by only 64% in thebaseline period and had an average of 13 mg/L present in the digester(Table 6).

Further evidence of complete nitrification and denitrification isillustrated by the gain in alkalinity in the digester where the dailyaverage of alkalinity increased by 71% during the test period over thatpresent during the baseline period. Table 7 shows the digesteralkalinity.

TABLE 7 Analytical Results of Digester Alkalinity. Digester Alkalinity(mg/l) Baseline 304 Delivered 520 Microorganisms

Bleach or Chlorine Reduction

The test plant uses chlorine bleach to disinfect the wastewater effluentfrom the treatment plant. The chlorine also destroys any ammonia in theeffluent. The use of delivered microorganisms results in a significantdecrease in the amount of chlorine bleach being used. Table 8 showsgallons of chlorine bleach used during the testing period and during thebaseline period.

FIG. 3 shows the baseline average bleach use and the test period bleachuse. The use of the delivered microorganism decreased the average bleachused from 447 gallons per day to 298 gallons per day. Table 9 shows thedaily average mass use of bleach and bisulfite and the daily averagechlorine residual. The use of chlorine bleach requires the plant toemploy sulfur dioxide to reduce the residual chlorine level in theeffluent.

TABLE 8 Daily Average Chlorine Bleach Use Bleach Daily avg, gals Period(gals) # days bleach Baseline 28,580 64 447 Delivered 32,180 108 298Microorganisms

TABLE 9 Daily Average Chlorine Bleach Mass and Bisulfite and ResidualChlorine Levels CL₂ Period Bleach (Mt) SO2 (Mt) SO2/Bleach residualBaseline 2.17 0.124 0.057 4.742 Delivered 1.35 0.123 0.091 4.579Microorganisms

Sour Reduction

The performance of the delivered microorganism shows a reduction inmixed liquor SOUR measured as milligram of oxygen per liter per gram VSSper hour. Samples are taken from the aeration basin and analyzed bystandard thermogravimetric analysis (TGA) following EPA method 160.2.Typical values for samples from an aeration basin fall in a range fromabout 15 to 20 mg O₂/L/g VSS/hr. SOUR values decreased to a lower rangeafter application of the delivered microorganism to values ranging from3 to 6 mg O₂/L/g VSS/hr.

FIG. 4 shows data from the baseline time period, recognized standardvalues, and data from the testing period when delivered microorganismwere used. The SOUR data measured during the testing period fall wellbelow the typical values, which are shown as a solid line in FIG. 4 .

Example 2: Phosphorous Removal

A test facility was selected at the wastewater treatment plant inBeaumont, Tex. This specific plant had an excellent record of meetingcompliance. The wastewater treatment facility had an average daily flowrate of 20 million gallons per day wastewater treated. Testing began inSpring 2016 with the introduction of microorganisms loaded onto aninorganic porous medium, or delivered microorganisms, to a Trickling BedFilter plant at a rate of 30 pounds per day. See FIG. 2 for a diagram ofa treatment system, microorganism delivery locations, and samplinglocations. The test period covered a total of four months, from April2016 to June 2016.

Orthophosphate levels in the influent are tracked along with phosphatelevels leaving the secondary clarifiers (SCE) the day following theaddition of the delivered microorganism. Orthophosphate levels andphosphate levels were analyzed using Standard Methods EPA approved4500-P phosphorus analysis. The orthophosphate and phosphate levels werecompared with phosphorous levels in the large retention pond two daysafter adding the delivered microorganisms. FIG. 5 shows theorthophosphate level of the wastewater leaving the SCE. Open circlesrepresent data taken during the testing period and the dashed linerepresents the linear regression of the data. The data show a slightincreasing trend in phosphate removal from the entering raw sewage toafter SCE treatment. FIG. 6 shows the orthophosphate level of wastewaterin the large retention pond. Open circles represent data taken duringthe testing period and the dashed line represents the linear regressionof the data. The data shows a steep increasing trend in phosphateremoval from entering raw sewage to wastewater in the large retentionpond.

Example 3: Zeolite Use

A series of experiments utilizing zeolite as the inorganic porous mediumwas analyzed for microbial growth and substrate uptake. The zeolite usedwas Clinoptilolite, Natural Zeolite, CAS number 12173-10-3. The controlwas a liquid based mixed microbial culture. The zeolite was loaded withthe exact quantities of the mixed cultures. FIG. 7 shows the total massof gas produced for the liquid control and for the microorganisms loadedonto zeolite. The zeolite containing culture shows an increase in totalmass of gas produced compared to the control. FIG. 8 shows the acidproduction for both the control and zeolite containing culture. Thezeolite containing culture shows an increase in acid produced comparedto the control. FIG. 9 shows the sugar uptake for both the control andthe zeolite containing culture. The zeolite containing culture shows anincrease in sugar uptake compared to the control.

1.-19. (canceled)
 20. An assemblage for treating wastewater andproducing fertilizer or compost, the assemblage comprising: a reactionvessel comprising one or more inlets configured to receive an influentstream and a microbial solution, and one or more outlets to direct aneffluent stream out of the reaction vessel, wherein the influent streamcomprises an aqueous phase and a biosolids phase, and wherein themicrobial solution comprises at least one microbial species loaded ontoor within an inorganic porous medium; and a separator comprising atleast one inlet stream and at least one outlet stream, wherein theseparator is configured to separate the effluent stream into a treatedaqueous phase and a treated biosolids phase.
 21. The assemblage of claim20, wherein the inorganic porous medium comprises silica, zeolite,aluminosilicate, silicate, diatomaceous earth, or any combinationthereof.
 22. The assemblage of claim 20, wherein the biosolids phasecomprises at least one nutrient source for the at least one microbialspecies.
 23. The assemblage of claim 20, wherein the reaction vessel isconfigured to operate under anaerobic condition.
 24. The assemblage ofclaim 20, wherein the reaction vessel is configured to operate underaerobic condition.
 25. The assemblage of claim 20, wherein the at leastone microbial species comprises a facultative microbial species.
 26. Theassemblage of claim 20, wherein the reaction vessel is configured tocycle through aerobic and anaerobic conditions.
 27. The assemblage ofclaim 20, wherein the inorganic porous medium comprises a physicalmodification configured to aggregate or agglomerate the inorganic porousmedium.
 28. The assemblage of claim 27, wherein the physicalmodification comprises a roughened surface.
 29. The assemblage of claim20, wherein the inorganic porous medium comprises a chemicalmodification configured to precipitate or preferentially bind ammoniaand phosphorous.
 30. The assemblage of claim 29, wherein the chemicalmodification comprises a chelating agent.
 31. The assemblage of claim29, wherein the chemical modification comprises a salt.
 32. Theassemblage of claim 29, wherein the chemical modification comprises aligand.
 33. The assemblage of claim 20, wherein the influent streamcomprises residential wastewater, agricultural wastewater, industrialwastewater, runoff wastewater, or any combination thereof.
 34. Theassemblage of claim 20, wherein the separator comprises a decanter,filter, centrifuge, or any combination thereof.
 35. The assemblage ofclaim 20, wherein the reaction vessel is configured to be operated in astationary phase such that a number of the at least one microbialspecies produced is equal to a number of microbial species consumed andan overall population of the at least one microbial species remainsunchanged.
 36. The assemblage of claim 20, further comprising a returnactivated sludge (RAS) line that carries a portion of the treatedbiosolids phase from the separator to the reaction vessel.
 37. Theassemblage of claim 20, further comprising a waste activated sludge(WAS) line that carries a portion of the treated biosolids phase fromthe separator to a dewaterer configured to dewater the treated biosolidsphase to produce the fertilizer or compost.
 38. The assemblage of claim20, wherein the reaction vessel comprises an anaerobic digester.